CN104622907A - Use of starfish extract for preparing composition for promoting collagen proliferation - Google Patents

Use of starfish extract for preparing composition for promoting collagen proliferation Download PDF

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Publication number
CN104622907A
CN104622907A CN201410625151.6A CN201410625151A CN104622907A CN 104622907 A CN104622907 A CN 104622907A CN 201410625151 A CN201410625151 A CN 201410625151A CN 104622907 A CN104622907 A CN 104622907A
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extract
asterias amurensis
amurensis lutken
collagen
purposes
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CN201410625151.6A
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Chinese (zh)
Inventor
苏香绫
林咏翔
林咏凯
陈盈如
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BAIYUETE COSMETICS (SHANGHAI) CO LTD
Shanghai Biotech Co ltd
BAIYUETE BIOTECHNOLOGY (SHANGHAI) CO LTD
TCI Co Ltd
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BAIYUETE COSMETICS (SHANGHAI) CO LTD
Shanghai Biotech Co ltd
BAIYUETE BIOTECHNOLOGY (SHANGHAI) CO LTD
TCI Co Ltd
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Publication of CN104622907A publication Critical patent/CN104622907A/en
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  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)

Abstract

The invention discloses an application of starfish extract in preparing a composition for promoting collagen proliferation. The starfish extract contains various active ingredients, can effectively improve the collagen production capacity of skin fibroblasts, and the extraction method can effectively improve the economic value of the starfish and has no pollution, so the starfish extract can be effectively applied to skin care products and cosmetics.

Description

Asterias amurensis Lutken extract is for the preparation of the purposes of the compositions of promotion collagen hyperplasia
Technical field
The present invention relates to a kind of purposes promoting collagen hyperplasia, particularly a kind of Asterias amurensis Lutken extract promotes the purposes of the compositions of collagen hyperplasia in preparation.
Background technology
Asterias amurensis Lutken is the title of the animal of a guiding principle under Echinodermata, has surprising regeneration capacity, can not only bear the peduncle broken again, and some Asterias amurensis Lutken even can own regeneration one is complete by the peduncle be truncated new individuality.Asterias amurensis Lutken is distributed in the coastal area in Asia, North America, Australia more, and habitat is generally the seabed of husky mud substrate.Taiwan belongs to island, and Aquatic product enriches, and also has multiple Asterias amurensis Lutken, such as common in the Intertidal zone in Penghu style of calligraphy characterized by hollow strokes maple Asterias amurensis Lutken (Archaster typicus).Asterias amurensis Lutken of many uses, some Asterias amurensis Lutken, except can being provided as the material of food materials or processed food, is also a kind of often by the Chinese crude drug used; Asterias amurensis Lutken, because its high nutritive value, is also processed to mash feed.In medical usage, Asterias amurensis Lutken can be used to make harmless alternative blood plasma after refining; In addition, there is research to point out, the compositions such as the EPA contained by Asterias amurensis Lutken, to the treatment of hyperlipidemia, cardiovascular disease, have certain effect.
Collagen protein is a kind of biological polymer substance, is the protein that vertebrates in-vivo content is maximum, in human body, then account for 1/3rd of gross protein.Mainly being present in the extracellular matrix (extracellular matrix) of connective tissue, is cell and intercellular support.The skin, cartilage, cornea, blood vessel wall, internal organs etc. of human body are all containing collagen protein, and especially skin, collagen protein provides support the intensity of Skin Cell, keeps compacting and elasticity of skin.But with age and the accumulation of suffered external unfavorable factor, the ability that dermal fibroblasts generates collagen protein can reduce, once the speed that collagen protein runs off is greater than the speed that it generates, skin just can produce wrinkle, the phenomenon such as lax.
Therefore, the collagen protein generative capacity of replenishing collagen or lifting cell is the important topic of skin care.Be no lack of the product of replenishing collagen on the market, comprise modes such as smearing with external, inject or eat.But the molecule of collagen protein is large, the not easily horny layer of transdermal, the skin corium dermal fibroblasts that mode of smearing almost cannot make collagen protein arrive it to act on; If with the direct replenishing collagen of mode of injection, although the microgroove of skin can be reduced immediately, but the ferment that this injection enters in the collagen protein meeting body in body decomposes, need regular injection, process is complicated and cost is high, and injectable collagen albumen may produce immunological rejection, increase the risk on using.If carry out replenishing collagen with the edible food being rich in collagen protein, after collagen protein enters the intestines and stomach, structure i.e. destroyed formation of its triple helix wins peptide more, and many victory peptides cut into micromolecule via Digestive system, finally resolve into amino acid again.Health no doubt can utilize these amino acid synthetic proteins, but is not limited to collagen protein, therefore the limited efficiency of actual replenishing collagen.
In view of this, the collagen protein generative capacity system promoting human dermal fibroblast blast cell comparatively replenishing collagen more can guarantee effectively produce and utilize collagen protein, but there is no a kind of safe and effective product on the market at present, can promote the generation of collagen protein.
Summary of the invention
Edge this, one of the present invention object is to provide a kind of Asterias amurensis Lutken extract the promotes collagen hyperplasia purposes of compositions in preparation, and wherein this Asterias amurensis Lutken extract comprises water solublity polysaccharides, and this water solublity polysaccharides is Asterias amurensis Lutken polysaccharide.This Asterias amurensis Lutken extract can promote the collagen hyperplasia rate of cell, and this cell is dermal fibroblasts and comes from the mankind; Asterias amurensis Lutken extract of the present invention can promote that the collagen hyperplasia rate of this dermal fibroblasts is more than 80%.
In one embodiment of this invention, this Asterias amurensis Lutken extract is extracted by Asterias amurensis Lutken live body and obtains, and is that Asterias amurensis Lutken live body is via made by freezing, pulverizing, high-temperature extraction and alcohol extraction.
In addition, the compositions prepared by Asterias amurensis Lutken extract of the present invention can be externally applied liniment or external pharmaceuticals or external skin care products or cosmetics.It should be noted that, described externally applied liniment or external pharmaceuticals include but not limited to ointment or cream or adhesive preparation or attach stype or the artificial organ containing active component.In addition, described external skin care products or cosmetics can include but not limited to astringent or cosmetic cream or emulsion or foundation cream or foundation emulsion or sunscreen or bath shampoo.
Another object of the present invention is to provide a kind of promoter promoting hyperplasia collagen protein, and it comprises Asterias amurensis Lutken extract, and wherein this cell is dermal fibroblasts, and this Asterias amurensis Lutken extract comprises the water solublity polysaccharides of more than 89% (w/w).
Asterias amurensis Lutken extract of the present invention contains active component, and can promote that dermal fibroblasts generates collagen protein significantly.Asterias amurensis Lutken extract of the present invention, the Asterias amurensis Lutken extract source that it comprises is natural, and preparation method is simple, pollution-free, therefore effectively can be applied to product and the cosmetics of maintenance skin.
Embodiments of the present invention are further illustrated below by graphic for cooperation; following cited embodiment illustrates invention Speciality and application of the present invention; but not to limit scope of the present invention; anyly have the knack of this those skilled in the art; without departing from the spirit and scope of the present invention; when doing a little change and retouching, therefore protection scope of the present invention is when being as the criterion depending on the scope described in accompanying claims.
Accompanying drawing explanation
Fig. 1 represents the extraction procedure figure of Asterias amurensis Lutken extract of the present invention.
Fig. 2 represents that Asterias amurensis Lutken extract of the present invention promotes collagen hyperplasia ability.
Fig. 3 represents that Asterias amurensis Lutken extract of the present invention promotes that dermal fibroblasts generates the fluorescence staining microscope figure of collagen protein, wherein (a) control group; B () is with the Asterias amurensis Lutken extract process of 5% (v/v); C () processes with epidermal growth factor (EGF, 5ng/mL).
Detailed description of the invention
For confirming that Asterias amurensis Lutken extract of the present invention can promote collagen hyperplasia, the present invention be by gentle, operate easy method and prepare Asterias amurensis Lutken extract, and analyze the active component in Asterias amurensis Lutken extract.In addition, also by Asterias amurensis Lutken extract of the present invention and dermal fibroblasts co-cultivation, assess the excellent generative capacity of collagen protein that Asterias amurensis Lutken extract of the present invention promotes dermal fibroblasts.
Definition
Asterias amurensis Lutken extract of the present invention is the collagen protein production rate that can promote dermal fibroblasts, and the content being enough to realize this object is defined as " effective dose ".The factors known according to those who are familiar with this art decides by the content that can realize this object, the effect of the body weight of such as consumer, general body state, age, sex and constituent substrate.
Embodiment 1: the preparation of Asterias amurensis Lutken extract
Style of calligraphy characterized by hollow strokes maple Asterias amurensis Lutken (Archaster typicus) used in the present invention is collected from area, Penghu, Taiwan.About the preparation of Asterias amurensis Lutken extract, please refer to Fig. 1.In time preparing Asterias amurensis Lutken extract of the present invention, first, collect Asterias amurensis Lutken live body and in time catching, be placed in-4 DEG C ~-20 DEG C environment stored frozen immediately, by freezing Asterias amurensis Lutken live body grinding (as step S1); Then, by through grinding Asterias amurensis Lutken with 1:3 ~ 1:20 (w:v) ratio water-soluble and be heated to 90 DEG C ~ 100 DEG C extraction 1 ~ 5 hour (as step S2); After high-temperature extraction, filter with centrifugal action (4,300g) and get its supernatant (as step S3); Then, the ethanol adding 3 times of volumes in 4 DEG C makes polysaccharides precipitate and maintains 16-24 hour (as step S4); After alcohol settling effect, filter with centrifugal action (4,300g rpm) and get its precipitate (as step S5); Then, this precipitate is dissolved in (as step S6) in the water of 10-15 times of volume; Finally, filter with centrifugal action (4,300g rpm) and get its supernatant (as step S7), namely obtaining Asterias amurensis Lutken extract of the present invention.
Embodiment 2: Asterias amurensis Lutken extract water solublity polysaccharides is analyzed
Total polysaccharides content of Asterias amurensis Lutken extract of the present invention measures with phenol-sulphuric acid (phenol-sulfuric acid) method.The Glucose standards solution of 100 μ L Asterias amurensis Lutken extracts and variable concentrations (0,25,50,75,100 and 125mg/mL) is mixed in test tube.Then 0.5mL phenol solution is added and 2.5mL sulphuric acid reacts.Test tube is measured its light absorption value at 750nm with spectrum signal transducer (spectrophotometer).
Analysis result shows, water solublity polysaccharides containing 2.23% (w/v) in the Asterias amurensis Lutken extract solution of 2.5% (w/v), the water solublity polysaccharides content therefore contained by known Asterias amurensis Lutken extract of the present invention is approximately 89% (w/w).
Embodiment 3: Asterias amurensis Lutken extract promotes collagen hyperplasia capability analysis
Human skin fibroblast CCD-966SK (Human skin fibroblast cells is cultivated with the minimal essential medium (MEM) containing 10% hyclone (FBS), purchased from Foodstuff Industrial and Development Inst.) in 24-porose disc, the concentration of human skin fibroblast is 4 × 10 4cells/well.Culture medium is upgraded after 24 hours cultivate.Experimental group is dilute Asterias amurensis Lutken extract (concentration be respectively 0.2,0.4,0.6,0.8 and 1.0mg/mL) with phosphate buffered solution (phosphate buffer saline, PBS) and add in each hole.Control group does not then add Asterias amurensis Lutken extract, and in 37 DEG C, cultivate 48 hours under 5% carbon dioxide conditions after get its supernatant, experiment find Asterias amurensis Lutken extract promote collagen protein be created on 48 hours within to increase in time and linear relevant.
Use collagen Assay Kit (Bicolor Life Science Assays, Northern Ireland, UK) analyzes the content of water solublity I-type collagen.At room temperature, by supernatant 100 μ L and the 1mL of said method gained stain mixes 30 minutes.Then with 15,000g centrifugal action 10 minutes to form collagen deposit thing.Remove centrifugal after supernatant and collagen deposit thing is dissolved in 1mL basic solvent.Its light absorption value at wavelength 540nm is measured with spectrum signal transducer.The quantity of collagen protein calculates with water-solubility collagen standard curve (standard without disease Corii Bovis seu Bubali skin I-type collagen, the U.S.).
Please refer to Fig. 2, this figure is that Asterias amurensis Lutken extract of the present invention promotes the analysis that collagen protein generates, and find that experimental group is with after the Asterias amurensis Lutken extract process of 1mg/mL, per unit human skin fibroblast generates the collagen protein of 45.25 ρ g; Far above the amount generating 24.48 ρ g collagen protein in control group, therefore, Asterias amurensis Lutken extract of the present invention obviously confirms effectively to promote that collagen hyperplasia rate is more than at least 80%, goodly reaches 84.8%.
Embodiment 4: Asterias amurensis Lutken extract promotes the fluorescence immunoassay of collagen hyperplasia
Fix human skin fibroblast after CCD-966SK30 minute with the phosphate buffered solution (PBS) containing 1% paraformaldehyde (para-formaldehyde, Sigma, France) in addition, and rinse in phosphate buffered solution.This cell 24 hours are processed respectively with Asterias amurensis Lutken extract of the present invention (5% (v/v)), epidermal growth factor (5ng/mL), control group cell then processes with normal saline solution (Phosphate Buffered Saline, PBS).(1/2000 dilutes for the cell this distinctly processed through Asterias amurensis Lutken extract or epidermal growth factor again and water resistant dissolubility I-type collagen rat monoclonal antibody, abcam, Britain) co-cultivation is in the phosphate buffered solution containing 1% (w/v) bovine serum albumin (PBS).Then, clean up hill and dale with phosphate buffered solution and with the anti-rat IgG antibody of fluorescein (fluorescein) labelling (1/200 dilutes, KPL, the U.S.) co-cultivation 60 minutes under no light conditions.After phosphate buffered solution cleaning, observe with fluorescence conjugated focusing microscope (Leica TSC SP2, Germany).
Please refer to Fig. 3, this figure is the fluorescence staining microscope figure that dermal fibroblasts generates collagen protein, wherein (a) represents control group cell, b () representative is with the cell of 5% (v/v) Asterias amurensis Lutken extract process, (c) represents the cell processed with epidermal growth factor (5 ng/mL).In figure, arrow represents collagen protein.Clearly visible by Fig. 3, through the cell of Asterias amurensis Lutken extract of the present invention process, the growing amount of collagen protein is significantly increased compared with control group cell, also compared with the more collagen protein of the Hemapoiesis of epidermal growth factor process, therefore, Asterias amurensis Lutken extract of the present invention confirms the hypertrophy that effectively and significantly can promote collagen protein further.
Meanwhile, Asterias amurensis Lutken extract of the present invention also comprises multiple amino acid and trace element, and analysis result please refer to following table.
The amino acid content of table 1, Asterias amurensis Lutken extract
Amino acid Content (mg/g)
Hydroxyl proline Hydroxyproline 5.08
Aminothiopropionic acid Cystine 4.27
Histidine Histidine 5.39
Glutamic acid Glutamic acid 54.08
Bright amino acid Leucine 24.19
Alanine Alanine 21.7
Rely amino acid Lysine 26.07
This amino acid Serine 18.67
Different bright amino acid Isoleucine 15.18
Phenylalanine Phenylalanine 13.87
Soviet Union amino acid Threonine 17.69
Aspartic acid Aspartic acid 37.11
Pick amino acid Valine 16.77
Tryptophan Tryptophan 5.81
Proline Proline 17.31
Taurine Taurine 12.35
MET Methylthianine 8.69
Spermine acid Arginine 39.99
Glycine Glycine 33.71
The micronutrient levels of table 2, Asterias amurensis Lutken extract
Trace element Content (μ g/g)
Copper Cu 5.6
Plumbous Pb <0.2
Manganese Mn 74.5
Zinc Zn 77.6
Nickel 6.05
Calcium Ca 153000
Magnesium Mg 533
Phosphorus P 125
Cobalt Co 1.08
Selenium Se 2.16
Chromium Cr 1.40
Cadmium Cd 28.30
Silicon Si 5.30
Germanium Ge 5.90
Strontium Sr 1933
Embodiment 5: produce the cosmetics and/or pharmaceuticals that promote collagen hyperplasia with Asterias amurensis Lutken extract
Cosmetics and/or the pharmaceuticals such as Water-In-Oil (O/W) neutral cream and gel can be produced with Asterias amurensis Lutken extract.The neutral cream oil phase about about 30 ~ 50% of O/W, do not have too greasy impression, Asterias amurensis Lutken extract can be added in formula, there is the effect promoting skin collagen hypertrophy, by as shown in table 3 for the formula that the Asterias amurensis Lutken extract of gained of the present invention is applied to neutral cream:
Table 3, formula containing Asterias amurensis Lutken extract cream
Oil content: Octadecanol 6.0%
Stearic acid 2.0%
Agnolin 4.0%
Shark alkane 9.0%
Octyl group 12 alkanol 10.0%
Wetting agent PEG 1500 4.0%
1,3-fourth dioxane 6.0%
Interfacial agent Glyceryl monostearate 2.0%
POE (25) spermaceti alcohol ester 3.0%
Antiseptic 0.01%
Antioxidant 0.01%
Spice 0.02%
Active component Asterias amurensis Lutken extract 0.1%
Pure water 53%
Slowly add the composition of oil phase at high speeds, to be emulsified complete after, containing the cream of Asterias amurensis Lutken extract, this cream at room temperature can preserve 6 months.
The Asterias amurensis Lutken extract of gained of the present invention also can be used for producing collagen gel, and this gel also can reach the effect promoting skin collagen hypertrophy, and the formula of this gel is as shown in table 4:
Table 4, formula containing Asterias amurensis Lutken extract gel
Wetting agent Dipropylene glycol 7.0%
PEG 1500 5.0%
Water soluble polymer Methylcellulose 0.2%
Interfacial agent POE(15)oleyl alcohol ether 1.0%
Alkaline agent Sodium hydroxide 0.1%
Antiseptic 0.01%
Antioxidant 0.01%
Spice 0.02%
Active component Asterias amurensis Lutken extract 0.1%
Pure water 83%
Embodiment 6: produce the essence promoting collagen hyperplasia with Asterias amurensis Lutken extract
Asterias amurensis Lutken extract of the present invention can be used for producing the essence promoting collagen hyperplasia, comprises an Asterias amurensis Lutken extract component, a moisture-keeping functions composition component, a whitening function composition component and an antibacterial.The essence of promotion collagen hyperplasia is using deep seawater as substrate, and its content is in fact about 40wt%, wherein, the total weight of the essence of this promotion collagen hyperplasia, the content of this Asterias amurensis Lutken extract component is between 15wt% to 20wt%; This moisture-keeping functions composition component includes but not limited to be selected from the group be made up of glucoprotein (Glygoprotein), hyaluronic acid, 1,3 butylene glycol or 1,3-PD, and its content is between 10wt% to 20wt%; This whitening function composition component includes but not limited to be selected from the group be made up of Folium Vaccinii vitis-idaeae foline (Arbutin), kojic acid (Kojic Acid), vitamin C sodium phosphate (Sodium Ascorbyl Phosphate) or Magnesium Ascorbyl Phosphate (Magnesium Ascorbyl Phosphate), and its content is between 10wt% to 20wt%; And the content of this antibacterial is in fact 0.1wt% to 0.2wt%.
Asterias amurensis Lutken extract of the present invention is for the effect producing essence: in essence, Asterias amurensis Lutken extract component contains water solublity polysaccharide, i.e. Asterias amurensis Lutken polysaccharide, the collagen hyperplasia rate of dermal fibroblasts can be promoted, and also containing multiple amino acid and trace element in Asterias amurensis Lutken extract, can activating skin cells, moisture-keeping functions component in collocation essence and whitening function component, can promote essence maintenance effect to skin further.
In sum, Asterias amurensis Lutken extract of the present invention has various active composition, and be rich in water solublity polysaccharides, the collagen hyperplasia of dermal fibroblasts can be promoted, in addition, the manufacture method of Asterias amurensis Lutken extract of the present invention is simple and easy, pollution-free, therefore not only can effectively utilize natural resources and not to environmental pollution, also can provide a high activity and the skin care product raw material of safety.
Only, above describing is only preferred embodiment of the present invention and illustrates, is allly skillful in this those skilled in the art when can do other all improvement according to above-mentioned explanation, and only these changes still belong in spirit of the present invention and following defined the scope of the claims.

Claims (12)

1. Asterias amurensis Lutken extract promotes a purposes for the compositions of collagen hyperplasia in preparation, and wherein this Asterias amurensis Lutken extract comprises water solublity polysaccharides.
2. purposes as claimed in claim 1, wherein this Asterias amurensis Lutken extract can promote the collagen hyperplasia rate of cell.
3. purposes as claimed in claim 2, wherein this cell is dermal fibroblasts.
4. purposes as claimed in claim 3, wherein this dermal fibroblasts comes from the mankind.
5. the purposes as described in claim 3 or 4, wherein this Asterias amurensis Lutken extract promotes that the collagen hyperplasia rate of this dermal fibroblasts is more than 80%.
6. purposes as claimed in claim 1, wherein this water solublity polysaccharides is Asterias amurensis Lutken polysaccharide.
7. purposes as claimed in claim 1, wherein this Asterias amurensis Lutken extract is extracted by Asterias amurensis Lutken live body and obtains.
8. purposes as claimed in claim 1, wherein this Asterias amurensis Lutken extract is that Asterias amurensis Lutken live body is via made by freezing, pulverizing, high-temperature extraction and alcohol extraction.
9. purposes as claimed in claim 1, wherein said composition is externally applied liniment, external pharmaceuticals, external skin care products or cosmetics.
10. promote a promoter for hyperplasia collagen protein, comprise Asterias amurensis Lutken extract.
11. promoter as claimed in claim 10, wherein this cell is dermal fibroblasts.
12. promoter as claimed in claim 10, wherein this Asterias amurensis Lutken extract comprises the water solublity polysaccharides of more than 89% (w/w).
CN201410625151.6A 2013-11-14 2014-11-07 Use of starfish extract for preparing composition for promoting collagen proliferation Pending CN104622907A (en)

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KR101640799B1 (en) * 2016-03-11 2016-07-19 아무르콜라겐(주) Method for preparing starfish extract and cosmetic composition comprising starfish extract

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Application publication date: 20150520